ABCB5-positive stem cells for LSCD therapy

用于 LSCD 治疗的 ABCB5 阳性干细胞

基本信息

批准号：
10668673
负责人：
NATASHA Y FRANK
金额：
$ 80.11万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-01 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10668673
关键词：
Allogenic Autologous Bilateral Blindness Cell Surface Proteins Cell physiology Cells Chronic Clinical Clinical Trials Cornea Corneal Diseases Corneal Injury Corneal Stroma Development Epigenetic Process Epithelium Europe Eye FDA approved Funding Generations Genes Grant Homeostasis Human Immune Immunosuppressive Agents In Vitro Inflammation Ligands Molecular Monoclonal Antibodies Mus Natural regeneration Nature Organoids Pathway interactions Patients Prevention therapy Progress Reports Publications R24 Reporting Research Research Project Grants Residual state Signal Pathway Signal Transduction Signal Transduction Pathway Skin Source Stem cell transplant Surface Testing Therapeutic Therapeutic Trials Therapeutic immunosuppression Transplantation Vision adult stem cell allograft rejection c-myc Genes corneal epithelium corneal regeneration functional restoration human tissue in vivo induced pluripotent stem cell injured limbal manufacture neovascularization next generation novel strategies prevent programs repaired restoration self-renewal stem cell population stem cell proliferation stem cells success

项目摘要

ABSTRACT- limited to 30 lines The corneal epithelium is a rapidly self-renewing epithelial surface essential for maintaining a clear cornea and normal vision. The limbus contains a small subpopulation of limbal stem cells (LSCs) that continually repopulate the corneal epithelium and patients with limbal stem cell deficiency (LSCD) are unable to regenerate the corneal epithelium, resulting in "conjunctivalization" that ultimately leads to blindness. We previously identified that the ABCB5 gene is expressed by LSCs in both mouse and human tissues and that normal function of ABCB5+ LSCs is required for corneal development, homeostasis, and repair. Moreover, ABCB5 is a cell surface protein and specific monoclonal antibodies are capable of isolating pure ABCB5+ LSCs for study and transplantation. Research from our prior grants supported the first clinical trials using purified in vitro-expanded human allogeneic ABCB5+ LSCs for patients with bilateral LSCD (ClinicalTrials.gov: Identifier NCT03549299). Despite our success, the use of donor allogeneic ABCB5+ LSC transplants is suboptimal because it requires immunosuppressive treatment to prevent immune rejection of donor cells. Allogeneic donor ABCB5+ LSCs were used because LSCD patients lack a source of healthy autologous ABCB5+ LSCs since both eyes are involved. To solve this problem and to treat bilateral LSCD patients with autologous ABCB5+ LSCs, this grant renewal will study three novel approaches. The first approach uses transplantation of ABCB5+ iLSCs (induced LSCs derived from iPSCs generated from skin) using full reprogramming of autologous skin cells, using the Yamanaka genes OSKM (Oct4, Sox2, Klf4, c-Myc) to produce human induced pluripotent stem cells (iPSCs) that are then differentiated in vitro into 2D eye-like organoids that remarkably contain ABCB5+ LSCs (Watanabe et al., iScience, 2021). The second approach uses transplantation-independent restoration and expansion of residual dysfunctional or quantity-deficient LSCs, using in vivo partial epigenetic reprogramming, a new form of reprogramming discovered by Co-PI Dr. Bruce Ksander and collaborators (Lu et al., Nature, 2020), which uses transient expression of three Yamanaka genes, OSK, to restore and rescue injured dysfunctional cells in vivo. The third approach uses niche-produced LAMA5 ligand-based signaling activation of the BCAM molecular pathway that we just discovered is critical to ABCB5+ LSC and Transient Amplifying Cell (TAC) expansion (Sasamoto et al. Cell Reports, 2022, in press), with resultant stem cell proliferation programs that enhance corneal regeneration. Thus, our hypothesis is that it is possible to manufacture and expand autologous ABCB5+ LSCs from patients with bilateral LSCD using (i) full reprogramming in vitro, (ii) partial epigenetic reprogramming in vivo, and (iii) niche-produced LAMA5 ligand-based activation of LSCs through specific BCAM/ABCB5 signal transduction pathways. These autologous ABCB5+ LSC will fully regenerate and maintain a normal limbus and corneal epithelium. This hypothesis will be tested in three independent specific aims that are thematically related.

摘要限制为30行角膜上皮是一种快速自我更新的上皮表面，对于保持透明角膜和正常视力。边缘包含一个小小的缘干细胞（LSC）的小亚群重新填充角膜上皮和边缘干细胞缺乏症患者（LSCD）无法再生角膜上皮，导致“结膜化”最终导致失明。我们以前确定的是，ABCB5基因在小鼠和人体组织中均由LSC表达 ABCB5+ LSC的正常功能是角膜发育，稳态和修复所必需的。而且， ABCB5是一种细胞表面蛋白，特定的单克隆抗体能够分离纯ABCB5+ 研究和移植的LSC。我们先前的补助金的研究支持了首次使用的临床试验为双侧LSCD患者（ClinicalTrials.gov：标识符NCT03549299）。尽管我们成功了，但使用供体同种异体ABCB5+ LSC移植是次优的是因为它需要免疫抑制治疗以防止免疫排斥供体细胞。使用同种异体供体ABCB5+ LSC，因为LSCD患者缺乏健康自体的来源 ABCB5+ LSC由于两只眼睛都涉及。解决此问题并治疗双侧LSCD患者自体ABCB5+ LSC，此授予续签将研究三种新方法。第一种方法使用使用完整使用Yamanaka基因OSKM（OCT4，SOX2，KLF4，C-MYC）重新编程自体皮细胞生产人类诱导的多能干细胞（IPSC），然后在体外分化为2D眼样非常包含ABCB5+ LSC的类器官（Watanabe等，Iscience，2021）。第二种方法使用非依赖于移植的恢复和剩余功能失调或数量缺陷的扩展 LSC，使用体内部分表观遗传重编程，这是Co-Pi发现的一种新形式的重编程形式 Bruce Ksander博士和合作者（Lu等人，自然，2020年），使用三个的瞬态表达 Yamanaka基因，OSK，在体内恢复和营救受伤的功能失调的细胞。第三种方法使用生产利基生产的LAMA5配体基于BCAM分子途径的信号传导激活我们只是我们发现对ABCB5+ LSC和瞬时扩增细胞（TAC）膨胀至关重要（Sasamoto等。报道，2022年，印刷中），导致了增强角膜再生的干细胞增殖计划。因此，我们的假设是可以从患者中生产和扩展自体ABCB5+ LSC 使用（i）在体外进行完整重新编程的双边LSCD，（ii）体内部分表观遗传重编程和（iii）利基生产的LAMA5配体通过特定BCAM/ABCB5信号转导基于LSC的激活途径。这些自体ABCB5+ LSC将完全再生并保持正常的边缘和角膜上皮。该假设将以主题相关的三个独立特定目的进行检验。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Isolation and detection of circulating tumour cells from metastatic melanoma patients using a slanted spiral microfluidic device.

DOI：
10.18632/oncotarget.18641
发表时间：
2017-09-15
期刊：
Oncotarget
影响因子：
0
作者：
Aya-Bonilla CA;Marsavela G;Freeman JB;Lomma C;Frank MH;Khattak MA;Meniawy TM;Millward M;Warkiani ME;Gray ES;Ziman M
通讯作者：
Ziman M

[Stem cell-based strategies in vascular surgery].